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Human MLNR-FLAG Stable Cell Line-HEK293T

Cat.No.: CSC-RG0460
Cell Line Description: HEK293T-HuMLNR-FLAG cell line is a hypotriploid human cell line, which has been transfected with a human motilin receptor (MLNR) tagged in the N-terminus with FLAG to allow stably express of the human MLNR tagged in the N-terminus with FLAG. It is an example of a cell line transfected using our proprietary CBTGS gene screening and amplification system.
Background: Motilin is a 22-amino acid peptide hormone that is expressed throughout the gastrointestinal (GI) tract as well as in enteric neurons of the duodenum and colon. It plays an important role in gastric motility by stimulating interdigestive antrum and duodenal contractions. Development of motilin receptor agonists or antagonists could be useful in the treatment of GI disorders.
Growth Properties: Adherent
Morphology: Epithelial
Host Cell: HEK293T
Cell Line Validation: 1. Gene expression: RT-PCR experiments determined specific expression of human MLNR.2. Protein expression: MLNR expression in this cell line has been validated by WB.3. Functional validation: Functional assays.
Sub-type: Motilin
Propagation: Complete growth medium: DMEM, 10% FBS, 1 μg/mL puromycinAtmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37.0°C
Starting Cells From Frozen Cell Stock: 1. Remove the packaging cell lines from liquid nitrogen and carry out a quick thaw. Float the cells in the 37°C water bath for 2 minutes until nearly (80%) thawed. Once cells are thawed, it is important to dilute the cells 1:10 in growth media immediately to reduce the potentially toxic effects of the DMSO preservative on the cells.2. Clean the outside of the vial with 70% ethanol before opening.3. Remove the cells from the vial and add slowly into a 15ml conical tube containing 10 ml pre-warmed media. 4. Centrifuge for 3 minutes 1000 xg to pellet cells and remove the supernatant.5. Add 14 ml of media and transfer cells to a T25 flask or a 100 mm culture dish.6. Place the cells in the 37°C incubator with 5% CO2.7. Allow incubation for 3-4 days to reach confluence. The cells will re-attach to the surface over a period of several days in culture at 37°C.
Subculturing: 1. Remove and discard culture medium.2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.3. Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.4. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.5. Add appropriate aliquots of the cell suspension to new culture vessels.Incubate cultures at 37°C.Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended Medium Renewal: 2 to 3 times per week
Mycoplasma: Mycoplasma Status: Negative (MycoAlert Kit)
Freeze Medium: Complete growth medium 90%; DMSO, 10%
Storage: Liquid nitrogen
Preservation: 1. Detach cells from culture dish according to the Sub-Culture Procedure.2. Resuspend cells at a density of 5 x 10^6 cells/mL in freeze medium.Note: A T-75 culture flask typically yields enough cells for preparing two frozen vials.3. Aliquot 1 mL cells into cryogenic vials.4. Place vials in a freezing container and store at –80 °C overnight.5. Transfer vials to liquid nitrogen for long term storage. If properly stored, cells should remain stable for years.
Safety Considerations: The following safety precautions should be observed.1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum.2. No eating, drinking or smoking while handling the stable line.3. Wash hands after handling the stable line and before leaving the lab.4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells.5. All waste should be considered hazardous.6. Dispose of all liquid waste after each experiment and treat with bleach.
Ship: Dry ice
Gene Name: MLNR motilin receptor [ Homo sapiens ]
Official Symbol: MLNR
Synonyms: MLNR; GPR38; MTLR1; motilin receptor; G protein-coupled receptor 38
Gene ID: 2862
mRNA Refseq: NM_001507.1
Protein Refseq: NP_001498.1
MIM: 602885
UniProt ID: O43193
Chromosome Location: 13q14-q21
Pathway: Class A/1 (Rhodopsin-like receptors); G alpha (q) signalling events; GPCR downstream signaling; GPCR ligand binding; GPCRs, Class A Rhodopsin-like; Gastrin-CREB signalling pathway via PKC and MAPK; Neuroactive ligand-receptor interaction;
Function: G-protein coupled receptor activity; growth hormone-releasing hormone receptor activity
Products Types ◆ Recombinant Protein

◆ Assay kits

Research Area GPCR Proteins
Related Gene

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Optional requirements on this protein    +Expand
C-fusion    N-fusion   Non-tagged
His    GST   Fc   Others
<1.0 eu/μg    <0.1 eu/μg   <0.01 eu/μg   Not required
Monomer Isolation    Dimer Isolation    Not required
>80% by SDS-PAGE    >90% by SDS-PAGE   >95% by SDS-PAGE   Others

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